Conjugative transfer can be inhibited by blocking relaxase activity within recipient cells with intrabodies

Garcillán-Barcia, M. Pilar; Jurado, Paola; González-Pérez, Blanca; Moncalián, Gabriel; Fernández, Luis A.; Cruz, Fernando de la

doi:10.1111/j.1365-2958.2006.05523.x

Cited by 66 publications

(54 citation statements)

References 50 publications

(72 reference statements)

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“…Strains ␤2150 or BW27783 were used as donors, while DH5␣ was used as a recipient strain. Conjugation frequencies were expressed as the number of transconjugants per donor cell and calculated as described previously (21). Natural transformation.…”

Section: Methodsmentioning

confidence: 99%

Plasmid Conjugation from Proteobacteria as Evidence for the Origin of Xenologous Genes in Cyanobacteria

et al. 2014

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Comparative genomics have shown that 5% of Synechococcus elongatus PCC 7942 genes are of probable proteobacterial origin. To investigate the role of interphylum conjugation in cyanobacterial gene acquisition, we tested the ability of a set of prototype proteobacterial conjugative plasmids (RP4, pKM101, R388, R64, and F) to transfer DNA from Escherichia coli to S. elongatus. A series of BioBrick-compatible, mobilizable shuttle vectors was developed. These vectors were based on the putative origin of replication of the Synechococcus resident plasmid pANL. Not only broad-host-range plasmids, such as RP4 and R388, but also narrower-host-range plasmids, such as pKM101, all encoding MPF T -type IV secretion systems, were able to transfer plasmid DNA from E. coli to S. elongatus by conjugation. Neither MPF F nor MPF I could be used as interphylum DNA delivery agents. Reciprocally, pANL-derived cointegrates could be introduced in E. coli by electroporation, where they conferred a functional phenotype. These results suggest the existence of potentially ample channels of gene flow between proteobacteria and cyanobacteria and point to MPF T -based interphylum conjugation as a potential mechanism to explain the proteobacterial origin of a majority of S. elongatus xenologous genes.

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Section: Methodsmentioning

confidence: 99%

Plasmid Conjugation from Proteobacteria as Evidence for the Origin of Xenologous Genes in Cyanobacteria

et al. 2014

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“…It is also thought to energize the process of DNA transport (135,136). The conjugative mating channel is basically a protein secretion channel, which transports the relaxase protein bound to the DNA to be transferred (43,58). According to the nomenclature of protein secretion mechanisms, it is a T4SS (54).…”

Section: General Properties Of Plasmid Mobilitymentioning

confidence: 99%

Mobility of Plasmids

Smillie

Garcillán-Barcia

Francia

et al. 2010

Microbiol Mol Biol Rev

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955

1,054

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International audiencePlasmids are key vectors of horizontal gene transfer and essential genetic engineering tools. They code for genes involved in many aspects of microbial biology, including detoxication, virulence, ecological interactions, and antibiotic resistance. While many studies have decorticated the mechanisms of mobility in model plasmids, the identification and characterization of plasmid mobility from genome data are unexplored. By reviewing the available data and literature, we established a computational protocol to identify and classify conjugation and mobilization genetic modules in 1,730 plasmids. This allowed the accurate classification of proteobacterial conjugative or mobilizable systems in a combination of four mating pair formation and six relaxase families. The available evidence suggests that half of the plasmids are nonmobilizable and that half of the remaining plasmids are conjugative. Some conjugative systems are much more abundant than others and preferably associated with some clades or plasmid sizes. Most very large plasmids are nonmobilizable, with evidence of ongoing domestication into secondary chromosomes. The evolution of conjugation elements shows ancient divergence between mobility systems, with relaxases and type IV coupling proteins (T4CPs) often following separate paths from type IV secretion systems. Phylogenetic patterns of mobility proteins are consistent with the phylogeny of the host prokaryotes, suggesting that plasmid mobility is in general circumscribed within large clades. Our survey suggests the existence of unsuspected new relaxases in archaea and new conjugation systems in cyanobacteria and actinobacteria. Few genes, e.g., T4CPs, relaxases, and VirB4, are at the core of plasmid conjugation, and together with accessory genes, they have evolved into specific systems adapted to specific physiological and ecological contexts

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“…The relaxase also "pilots" the T strand through the translocation channel. In the recipient cell, the relaxase catalyzes the recircularization of the T strand and may also participate in second-strand synthesis or recombination into the chromosome (52,84,105).…”

Section: Conjugation Systemsmentioning

confidence: 99%

Biological Diversity of Prokaryotic Type IV Secretion Systems

Martı́nez

Christie

2009

Microbiol Mol Biol Rev

622

780

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SUMMARY Type IV secretion systems (T4SS) translocate DNA and protein substrates across prokaryotic cell envelopes generally by a mechanism requiring direct contact with a target cell. Three types of T4SS have been described: (i) conjugation systems, operationally defined as machines that translocate DNA substrates intercellularly by a contact-dependent process; (ii) effector translocator systems, functioning to deliver proteins or other macromolecules to eukaryotic target cells; and (iii) DNA release/uptake systems, which translocate DNA to or from the extracellular milieu. Studies of a few paradigmatic systems, notably the conjugation systems of plasmids F, R388, RP4, and pKM101 and the Agrobacterium tumefaciens VirB/VirD4 system, have supplied important insights into the structure, function, and mechanism of action of type IV secretion machines. Information on these systems is updated, with emphasis on recent exciting structural advances. An underappreciated feature of T4SS, most notably of the conjugation subfamily, is that they are widely distributed among many species of gram-negative and -positive bacteria, wall-less bacteria, and the Archaea. Conjugation-mediated lateral gene transfer has shaped the genomes of most if not all prokaryotes over evolutionary time and also contributed in the short term to the dissemination of antibiotic resistance and other virulence traits among medically important pathogens. How have these machines adapted to function across envelopes of distantly related microorganisms? A survey of T4SS functioning in phylogenetically diverse species highlights the biological complexity of these translocation systems and identifies common mechanistic themes as well as novel adaptations for specialized purposes relating to the modulation of the donor-target cell interaction.

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Conjugative transfer can be inhibited by blocking relaxase activity within recipient cells with intrabodies

Cited by 66 publications

References 50 publications

Plasmid Conjugation from Proteobacteria as Evidence for the Origin of Xenologous Genes in Cyanobacteria

Plasmid Conjugation from Proteobacteria as Evidence for the Origin of Xenologous Genes in Cyanobacteria

Mobility of Plasmids

Biological Diversity of Prokaryotic Type IV Secretion Systems

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